[Primary renal marginal zone B lymphoma]. / L'image du mois. Lymphome B primitif de la zone marginale rénale.

Primary renal marginal zone B lymphoma is extremely rare. The renal involvement by the lymphoma is most often a manifestation of a disseminated disease. We report the case of a 53-year-old patient who presented lower back pain and kidney failure. The renal biopsy showed a marginal zone B lymphoma type MALT and the PET scan showed bilateral renal involvement. A treatment based on rituximab combined with chemotherapy (cyclophosphamide, prednisone and oncovin) was started with good outcome.

Le lymphome B de la zone marginale rénale comme lésion primitive est extrêmement rare car l'atteinte rénale est le plus souvent une manifestation de la maladie disséminée. Nous rapportons le cas d'un patient âgé de 53 ans présentant des lombalgies chroniques avec insuffisance rénale. La biopsie rénale était en faveur d'un lymphome B de la zone marginale type MALT et la tomographie par émission de positons a objectivé une atteinte rénale bilatérale. Un traitement à base de rituximab couplé à une chimiothérapie (cyclophosphamide, prednisone et oncovin) a été démarré avec une bonne évolution.

Assessment of the effectiveness of wetland protection in improving waterbird diversity in a Moroccan wetland system.

Assessment of the effectiveness of nature conservation measures is a crucial step to ensure that the actions undertaken have achieved the expected results. Depending on the results of such evaluations, managers may need to reframe or even change their approaches. In this analysis, the effectiveness of establishing strictly protected areas was assessed in three important Moroccan Atlantic sites, namely Ad-Dakhla bay, Khnifiss lagoon, and Souss estuary. In particular, we investigated whether protection of these sites was followed by significant changes in the direction and/or strength of change in waterbird species richness and density over time. Our results showed that during the pre-protection period, waterbird species richness has been declining over time, while total waterbird density and the diversity of wading and open-water birds had been maintained at low levels. However, shortly after site protection, a rapid increase in these measures was observed, while there was no significant change in waterfowl species richness and density. Overall, these results suggest that the protection of these wetlands has been a watershed in the recent history of their birds, reversing the trends of a continuing decline towards rapid regeneration. This is one obvious signs of the success of the programme undertaken by the Moroccan authorities to conserve the biodiversity of these wetlands. Further investigations of the effects of protection at all wetlands throughout Morocco are needed to more profoundly understand the role of such conservation measures on shaping wintering waterbird communities.

A majority of parents of children with peanut allergy fear using the epinephrine auto-injector.

Prompt epinephrine administration is crucial in managing anaphylaxis, but epinephrine auto-injectors (EAIs) are underutilized by patients and their families. Children with peanut allergy were recruited from the Allergy Clinics at the Montreal Children's Hospital, food allergy advocacy organizations and organizations providing products to allergic individuals. Parents of children who had been prescribed an EAI were queried on whether they were fearful of using it and on factors that may contribute to fear. A majority of parents (672/1209 = 56%) expressed fear regarding the use of the EAI. Parents attributed the fear to hurting the child, using the EAI incorrectly or a bad outcome. Parents whose child had longer disease duration or a severe reaction and parents who were satisfied with the EAI training or found it easy to use were less likely to be afraid. Families may benefit from simulation training and more education on the recognition and management of anaphylaxis.

The alternative analog plasticizer BPS displays similar phenotypic and metabolomic responses to BPA in HepG2 and INS-1E cells.

Bisphenols A (BPA) and S (BPS) are endocrine-disrupting chemicals that affect energy metabolism, leading to impairment of glucose and lipid homeostasis. We aimed at identifying metabolic pathways regulated by both compounds in human liver cells and rat pancreatic ß-cells that could impair energy homeostasis regulation. We assessed the effects on growth, proliferation, and viability of hepatocarcinoma (HepG2) and insulinoma (INS-1E) cells exposed to either BPA or BPS in a full range concentration between 0.001 and 100 µM. Both the dose and duration of exposure caused a differential response on growth and viability of both cells. Effects were more pronounced on HepG2, as these cells exhibited non-linear dose-responses following exposure to xenobiotics. For INS-1E, effect was observed only at the highest concentration. In addition, we profiled their intracellular state by untargeted metabolomics at 24, 48, and 72 h of exposure. This analysis revealed time- and dose-dependently molecular changes for HepG2 and INS-1E that were similar between BPA and BPS. Both increased levels of inflammatory mediators, such as metabolites pertaining to linolenic and linoleic acid metabolic pathway. In summary, this study shows that BPS also disrupts molecular functions in cells that regulate energy homeostasis, displaying similar but less pronounced responses than BPA.

Antimicrobial susceptibility testing determined by Alfred 60/AST (Alifax®) in a multi-sites lab: performance's evaluation and optimization of workflow.

PURPOSE: New techniques are needed to speed-up the identification and antimicrobial susceptibility testing (AST) of bacteria associated with bloodstream infections. Alfred 60/AST (Alifax®, Polverara, Italy) performs AST by light scattering directly from positive blood cultures. METHODS: We evaluated Alfred 60/AST performances for 4 months. Each new episode of bacteraemia was included and AST were compared to either our rapid automated AST (Vitek® 2) or disk diffusion method. The discrepancies were investigated using Etest®. The time-to-result (TTR) was evaluated by comparing the blood volume inserted into Alfred 60/AST, i.e. 2 versus 7 blood drops. Taking into account the TTR, the workflow of positive blood cultures and the availability of AST results was studied in order to optimize the implementation of Alfred 60/AST. RESULTS: A total of 249 samples and 1108 antibiotics for AST were tested. After exclusion of unavailable results, 1008 antibiotics were analysed. 94.9% (n = 957/1008) of the antibiotics showed categorical agreement. There were 14 very major errors (VME), 24 major errors (ME) and 13 minor errors (mE). The VME were mostly related to clindamycin (64.3%) whereas meropenem and piperacillin-tazobactam constituted the major part (37.5% and 61.5%) of ME and mE respectively. Results were highly reliable for Enterobacterales and enterococci. The mean TTR ranged between 4.3 and 6.3 h and was statistically 20 min faster when applying the 7 blood drops protocol. We showed that Alfred 60/AST could give relievable results within working hours for positive blood culture which are flagged the same day between 12:00 am and 12:00 pm. CONCLUSION: Our study confirmed that Alfred 60/AST gives reliable AST results in a short period of time, especially for Enterobacterales and enterococci. AST could thus be easily obtained the same day of a positive blood culture. Clinical impact studies are mandatory to validate a 24/24 working.

A phospholipid-PEG2000 conjugate of a vascular endothelial growth factor receptor 2 (VEGFR2)-targeting heterodimer peptide for contrast-enhanced ultrasound imaging of angiogenesis.

The transition of a targeted ultrasound contrast agent from animal imaging to testing in clinical studies requires considerable chemical development. The nature of the construct changes from an agent that is chemically attached to microbubbles to one where the targeting group is coupled to a phospholipid, for direct incorporation to the bubble surface. We provide an efficient method to attach a heterodimeric peptide to a pegylated phospholipid and show that the resulting construct retains nanomolar affinity for its target, vascular endothelial growth factor receptor 2 (VEGFR2), for both the human (kinase insert domain-containing receptor - KDR) and the mouse (fetal liver kinase 1 - Flk-1) receptors. The purified phospholipid-PEG-peptide isolated from TFA-based eluents is not stable with respect to hydrolysis of the fatty ester moieties. This leads to the time-dependent formation of the lysophospholipid and the phosphoglycerylamide derived from the degradation of the product. Purification of the product using neutral eluent systems provides a stable product. Methods to prepare the lysophospholipid (hydrolysis product) are also included. Biacore binding data demonstrated the retention of binding of the lipopeptide to the KDR receptor. The phospholipid-PEG2000-peptide is smoothly incorporated into gas-filled microbubbles and provides imaging of angiogenesis in a rat tumor model.

Peripheral Medulloepithelioma: A Rare Entity to Know.

According to the World Health Organization, medulloepithelioma belongs to the embryonal neoplasm entity. It is a very rare, highly malignant tumor typically affecting infants and young children. Usually, the tumor arises in the eye or in the central nervous system; a peripheral location has been rarely reported without an established treatment. The recognition and separation of this neoplasm from other differential tumors are mandatory for better understanding of its biology and determination of optimal treatment. This paper reports a case of an ectopic intrapelvic medulloepithelioma with liver metastasis in a 3-year-old girl.

Outcome of Acute Myeloid Leukemia in Children Adolescents and Young Adults Treated with an Uniform Protocol in Casablanca, Morocco.

Treatment of acute myeloblastic leukemia in children, adolescents and young adults (AYA) is a challenge in low-income countries. To evaluate treatment outcomes of children (≤ 15 years) and AYA (15-30 years) diagnosed with novo AML and treated in a single center according to the AML-MA 2011 protocol. From January 2011 to December 2015, eligible patients (age ≤ 30 years) with novo AML had been enrolled on a uniform treatment protocol. The diagnosis was confirmed according to the FAB classification using the WHO 2008 criteria. Patients with WBC ≥ 50 G/L had pretreated 4 days of hydroxyurea followed by two inductions and two consolidations. Supportive care consisted of transfusion of labile blood products, antibiotics and antifungals, and patient and family education by the hygiene team. 155 patients were recruited, 41 were < 15 years old (22 boys, median age 7.8 years). Of the 114 AYA enrolled, (48 women, median age 23 years). Complete remission after two inductions was 28/41 (68.3%) of the children, including 100% of the children in the favorable group and 71/114 (62.3%) of the AYA, 22 of whom (68.7%) were in the favorable group. The number of deaths among children was 6 (14.6%). The evaluation of the AML-MA-2011 National Protocol in the age groups of children and AYA reveals that the objective of treatment is almost achieved in terms of complete remission in the two age groups.

Development and validation of a chemical hydrolysis method for dextromethorphan and dextrophan determination in urine samples: application to the assessment of CYP2D6 activity in fibromyalgia patients.

Dextromethorphan (DEM) is a widely used probe drug for human cytochrome P450 2D6 isozyme activity assessment by measuring the ratio between DEM and its N-demethylated metabolite dextrorphan (DOR). DOR is excreted in urine mainly conjugated to glucuronic acid. Prior to quantification, DOR must be deconjugated to avoid variability caused by the polymorphic glucuronosyltransferase enzyme. A chemical hydrolysis method was optimized using a chemometric approach. Three factors (acid concentration, hydrolysis time and temperature) were selected and simultaneously varied to study their effect on conjugated DOR hydrolysis. Hydrolysis conditions that maximize DOR release without significant degradation of both DEM and DOR were chosen and results were compared to those obtained by enzymatic method using beta-glucuronidase. An HPLC method with fluorescence detection was developed for the simultaneous quantitation of DEM, DOR and levallorphan, used as an internal standard. Separation was performed on a phenyl analytical column (150 mmx4.6 mm i.d., 5 microm) with a mobile phase consisting of 18% acetonitrile and 50 mM phosphoric acid (pH 3). The overall analytical procedure was validated and showed good performances in terms of selectivity, linearity, sensitivity, precision and accuracy. Finally, this assay was used to determine DEM/DOR molar ratios in fibromyalgia patients for the purpose of determining phenotype status for the CYP2D6.

[Pregnancy and delivery in a patient with Willebrand's disease. Apropos of a case]. / Maladie de von Willebrand et grossesse : à propos d'un cas.

Willebrand's disease is the most frequent inborn coagulopathy and type 3 is its most severe form. Pregnancy and delivery are critical events in women with Willebrand's disease of type 3. Prophylactic treatment for delivery and early postpartum period is recommended. We report the management of pregnancy and successful delivery of a 32-year-old woman with type 3. Prophylactic treatment with 2000 IU of Willebrand's disease factor (WdF) was given twice a day during the delivery day and the day after, and 1000 IU per day during the next three days. The patient did not show any spontaneous metrorrhagia but anemia. No bleeding was observed in the newborn.

[Role of Köllner-Hughes tarsoconjunctival flap in the reconstruction of large eyelid defects]. / Intérêt du lambeau tarsoconjonctival de Köllner-Hughes dans la reconstruction des larges défects palpébraux.

PURPOSE: To evaluate the aesthetic and functional results of the Köllner-Hughes tarsoconjonctival flap technique for large defects of over three-quarter of the lower eyelid. PATIENTS AND METHODS: A retrospective descriptive study of a series of 15 patients treated between January 2013 and December 2015. We included all patients who underwent reconstructive surgery for a tumor involving more than three-quarter of the lower eyelid and sparing the canthi. Reconstruction of the lower eyelid defect was performed using Köllner-Hughes technique. RESULTS: The mean age of our patients was 65.7±8 years. The sex ratio was 1.5 with male predominance. All patients had well-differentiated basal cell carcinoma of the lower eyelid. The average size of the eyelid defect induced by the surgery was 18±4mm. Release of the tarsoconjonctival flap was performed after a period of 8 to 10 weeks after the reconstructive surgery. After a mean follow-up of 12±6 months, the aesthetic and functional results were satisfactory in 86.7% of cases especially in terms of eyelid closure and protection of the ocular surface. Complications included erythema of the newly formed free edge in 26.7% of cases, keratinization and hypertrophy of the free edge in 20% of cases, the deformation of the upper free edge with an associated entropion in 20% of cases and lower eyelid retraction in 6.67% of cases. Ocular surface disease such assuperficial punctate keratitis or dry eye was found in 13.4% of cases. DISCUSSION: Several authors have chosen the Köllner-Hughes technique to treat increasingly wide eyelid defects with very satisfactory results. Eyelid reconstruction with the Köllner and Hughes technique is a very attractive procedure because it offers several advantages. It is a quick and easy technique that covers wide defects over three-quarter of the lower eyelid well with very satisfactory cosmetic results and without significantly increasing morbidity at the donor site. In combination with additional procedures, this technique can totally reconstruct the lower eyelid. Despite all its benefits, the Köllner-Hughes tarsoconjonctival flap has some minor drawbacks, such as the need for two surgeries spaced a few weeks apart to open the palpebral fissure; therefore it is contraindicated for one-eyed patients and children because of the risk of amblyopia. CONCLUSION: The tarsoconjonctival flap as initially described by Köllner and Hughes was indicated in the reconstruction of moderate and medium-sized defects of the lower eyelid. In our practice, we can extend the indications of this technique to much larger defects than three-quarter of the lower eyelid, while ensuring satisfactory aesthetic results and minimal complications.

[Anterior lamellar resection with lid margin split of the upper eyelid in the treatment of trachomatous entropion]. / Marginoplastie par recul de la lamelle antérieure dans le traitement de l'entropion trichiasis trachomateux de la paupière supérieure.

INTRODUCTION: The goal of this study is to assess functional and aesthetic results of anterior lamellar resection with lid margin splitting of the upper lid in the treatment of cicatricial trachomatous entropion. PATIENTS AND METHODS: Descriptive cross-sectional study of a series of 26 consecutive patients treated between January 2014 and December 2015. All patients were operated for cicatricial trachomatous entropion in our tertiary center using the technique of the anterior lamellar resection with lid margin splitting of the upper eyelid. All patients were followed for 6 to 12 months after surgery. The anatomical, functional and aesthetic results were evaluated six months after surgery. They were considered good if there was no recurrence of the entropion, no lashes in contact with the cornea and no associated eyelid complications. RESULTS: The mean age of the patients was 68.5±10 years with a male predominance (sex ratio=1.8). The average initial corrected visual acuity was 0.65±0.35 LogMAR, ranging from counting fingers at 1m to 6/10. Involvement was bilateral in 34.6% of cases. Correction of the cicatricial entropion was achieved in 24 patients (92.3% of cases) and full correction of misdirected lashes without any contact with the ocular surface was obtained in 23 patients (88.4% of cases). The lid margin was regular in 88.4% of cases. A significant improvement of the tear film and corneal surface was observed in 84.6% of patients. During the follow-up period, no cases of recurrent entropion were reported. DISCUSSION: Among the various surgical techniques, anterior lamellar resection with lid margin splitting is one that most respects the anatomy of the upper eyelid and allows precise intraoperative control of eyelid rotation and eversion of the misdirected lashes. Therefore, it reduces significantly the risk of recurrence and significantly enhances the aesthetic results of surgery. CONCLUSION: Anterior lamellar resection with lid margin splitting of the upper eyelid is a simple and effective technique that significantly improves the aesthetic result of cicatricial trachomatous entropion.

[Long-term results of phacoemulsification in pseudoexfoliation syndrome]. / Résultats à long terme de la phacoémulsification dans le syndrome pseudoexfoliatif.

INTRODUCTION: The prevalence of pseudoexfoliation syndrome is estimated at 8 % of all cataracts operated within our institution. The goal of our study was to describe the frequency of intraoperative and postoperative complications and long-term anatomical and functional results of phacoemulsification in pseudoexfoliation syndrome. PATIENTS AND METHODS: A retrospective observational study of a series including 103 patients (107 eyes) with cataract associated with pseudoexfoliation syndrome admitted between January 2010 and September 2014. All patients underwent phacoemulsification. RESULTS: The average age of our patients was 72.5±6 years. The average initial corrected visual acuity was 0.52±0.14LogMAR. The cataract was grade C3N3 in 58 % of cases. The pseudoexfoliation syndrome was bilateral in 69 % of cases. Pupillary dilatation was≤6mm in 54 % of cases. Among intraoperative complications, zonular dialysis was found in 1.8 % of cases. Posterior capsular rupture and vitreous loss was noted at 5.4 % of patients. The incidence of secondary cataract was 31.7 % at 12 months. The mean postoperative best corrected visual acuity was 0.13±0.15 LogMAR. DISCUSSION: The small pupil associated with high grade of the cataracts in our series explains the slightly higher frequency of intraoperative zonular dialysis. Cataract surgery by phacoemulsification allows for a statistically significant improvement in visual acuity (P<0.001). The incidence of secondary cataract, the main postoperative complication, is higher in our series compared to data in the literature. CONCLUSION: Pseudoexfoliation syndrome is common in our country. Functional results of phacoemulsification are very satisfactory, and secondary cataract is the main complication. A meticulous search for pseudoexfoliation is mandatory during the initial examination in order to reduce intraoperative complications. Careful removal of the cortex and systematic polishing of the lens capsule should limit the incidence of secondary cataracts.

Effects of nucleoside transport inhibition on hepatosplanchnic perfusion, oxygen extraction capabilities, and TNF release during acute endotoxic shock.

We explored the effects of the nucleoside transport inhibitor draflazine on regional blood flow, O2 extraction capabilities, and tumor necrosis factor (TNF) release in acute endotoxic shock. Fourteen anesthetized and mechanically ventilated dogs received 2 mg/kg of Escherichia coli endotoxin and were divided into two groups. Seven dogs received 0.1 mg/kg of draflazine 30 min before endotoxin, and 7 dogs served as a control group. Draflazine decreased arterial pressure without influencing cardiac index. Mesenteric and portal blood flow and ileum mucosal perfusion increased, but renal blood flow dramatically decreased. After endotoxemia, the draflazine-treated dogs had a lesser fall in cardiac index, filling pressures, and left ventricular stroke work index, and a lesser increase in pulmonary vascular resistance. After fluid resuscitation, they had a consistently lower renal blood flow and ileum mucosal perfusion, but a higher mixed venous and hepatic oxygen saturation and arterial pH than the control group. When cardiac index was reduced by tamponade to study the O2 extraction capabilities, renal blood flow and ileum mucosal perfusion remained lower in the draflazine group. Draflazine did not influence whole-body O2 extraction capabilities, but it delayed the occurrence of liver O2 supply dependency as indicated by a significantly lower liver DO2crit (27.7 +/- 3.9 vs. 43.3 +/- 10.8 mL/min) and a higher O2ERcrit (62.7 +/- 9.5 vs. 42.5 +/- 7.1%) than controls (both P< 0.05). On the other hand, draflazine increased intestinal DO2crit (42.4 +/- 15.4 vs. 27.7 +/- 6.5 mL/min, P < 0.05) compared to the control group. TNF levels remained higher in the draflazine group than in the control group, particularly 3 and 4 h after endotoxin administration. We conclude that nucleoside transport inhibition with draflazine does not alter global and hepatosplanchnic hemodynamics but may decrease gut mucosal perfusion and renal blood flow. However, this intervention can improve liver O2 extraction capabilities in acute endotoxic shock.

Simultaneous determination of scopolamine, hyoscyamine and littorine in plants and different hairy root clones of Hyoscyamus muticus by micellar electrokinetic chromatography.

Hyoscyamus muticus hairy root clones were established following infection with Agrobacterium rhizogenes strains A4, LBA-9402 and 15834 and with A. tumefaciens strain C58C1pRTGus104. The accumulation of tropane alkaloids hyoscyamine, littorine and scopolamine was evaluated by micellar electrokinetic capillary electrophoresis. Littorine was reported for the first time in these clones as well as in the roots of the intact plant and confirmed by collision induced dissociation-mass spectrometry. Tropane alkaloid content in hairy roots was compared with leaves and roots of normal plants at two vegetative stages. Significant differences appeared between the alkaloid contents of the different clones. In particular, all the hairy root clones and the roots of the intact plant produced 1.5-3 and 4.5-9 times more littorine than scopolamine, respectively. The only exception was clone KB7, carrying the h6h gene, which overproduced scopolamine. The aerial parts of H. muticus plants did not contain any littorine, thus indicating different transportation or translocation mechanisms of the various tropane alkaloids.

Calystegine distribution in some solanaceous species.

The distribution of eight calystegines (A(3), A(5), B(1), B(2), B(3), B(4), C(1) and N(1)) and their content was investigated by gas chromatography coupled to mass spectrometry (GC-MS) in Datura metel, Atropa belladonna, Hyoscyamus albus, Mandragora autumnalis, Solanum sodomaeum, Withania somnifera, Withania frutescens and Brunfelsia nitida. The most frequently encountered calystegines were A(3), B(1), B(2) and B(3), while distribution of N(1) and C(1) was more limited. In all the investigated samples, calystegines A(5) and B(4) were never detected. This report focuses for the first time on calystegines in Withania and Brunfelsia genera and in Mandragora autumnalis and Solanum sodomaeum species.

Development and robustness testing of a nonaqueous capillary electrophoresis method for the analysis of nonsteroidal anti-inflammatory drugs.

Nine non steroidal anti-inflammatory drugs were simultaneously separated by nonaqueous capillary electrophoresis with a methanol-acetonitrile (40:60, v/v) mixture containing 20 mM ammonium acetate. The effect of solvent composition, electrolyte nature and concentration on the electrophoretic behavior of the selected drugs was systematically studied. Investigated electrolytes were ammonium, lithium and sodium acetate. Modification of the solvent and/or the electrolyte composition was found to alter the migration order of the pharmaceutical drugs. Finally, to assess method robustness, three sensitive electrophoretic parameters as well as their interactions were evaluated using a full factorial design at two levels.

Simultaneous analysis of some amphetamine derivatives in urine by nonaqueous capillary electrophoresis coupled to electrospray ionization mass spectrometry.

A nonaqueous capillary electrophoresis method, coupled to UV and electrospray mass spectrometry (ESI-MS), is described for the simultaneous analysis of Ecstasy and other related derivatives. Several electrophoretic and ESI-MS parameters were systematically investigated, such as electrolyte nature and concentration, organic solvent and sheath liquid compositions, nebulization gas pressure and drying gas flow-rate. The best results were achieved with an acetonitrile-methanol (80:20, v/v) mixture containing 25 mM ammonium formate and 1 M formic acid, an applied voltage of 30 kV and a separation temperature of 15 degrees C. Under optimized CE-ESI-MS conditions, separation of the investigated drugs was performed in less than 6 min, with a high efficiency. Method precision based on migration time and peak area was determined and the limits of detection, which depend on the tested compound, were established between 20 and 70 ng ml(-1) in the selected ion monitoring mode. Finally, the described method was successfully applied to the analysis of amphetamines in urine after a liquid-liquid extraction.

Enantioseparation of atropine by capillary electrophoresis using sulfated beta-cyclodextrin: application to a plant extract.

A capillary zone electrophoresis (CZE) method, with sulfated beta-CD as chiral selector, was optimized by means of an experimental design for the enantioseparation of atropine. In this study, a central composite design was used and the following factors were varied simultaneously: buffer concentration, buffer pH and sulfated beta-CD concentration. The resolutions between littorine and its positional isomer ((-)-hyoscyamine) and between atropine enantiomers, as well as the separation time and generated current were established as responses. A model was obtained for each response by linear multiple regression of a second-degree mathematical expression. The most favorable conditions were determined by maximizing the resolution between atropine enantiomers and by setting the other responses at threshold values. Successful results were obtained with a 55 mM phosphate buffer at pH 7 in the presence of 2.9 mM sulfated-beta-CD at 20 degrees C and 20 kV. Under these optimized conditions, a baseline separation of littorine and atropine enantiomers was achieved in less than 5 min. Finally, the method allowed the enantiomeric separation of atropine in a pharmaceutical formulation and was also found to be suitable for the enantiomeric purity evaluation of (-)-hyoscyamine in plant extracts, in relation with the extraction procedure. It was demonstrated that supercritical fluid extraction induced less racemization than classical liquid-solid extraction procedures.

Simultaneous stereoselective analysis of tramadol and its main phase I metabolites by on-line capillary zone electrophoresis-electrospray ionization mass spectrometry.

On-line combination of partial filling capillary electrophoresis and electrospray ionization mass spectrometry was demonstrated for the simultaneous enantioseparation of tramadol and its main phase I metabolites. The partial filling technique was efficient at avoiding MS contamination by the chiral selector. Different experimental factors were investigated, including the chiral selector nature and concentration, plug length as well as the separation temperature. The best enantioseparation of the investigated compounds was achieved with a coated polyvinyl alcohol capillary and a 40 mM ammonium acetate buffer, pH 4.0, adding sulfobutyl ether beta-cyclodextrin (2.5 mg/ml) as the chiral selector. The charged cyclodextrin not only allowed enantioseparation of tramadol and its metabolites, but also improved the selectivity of compounds with the same molecular mass. Finally, CE-electrospray ionisation-MS was successfully applied to the stereoselective analysis of tramadol and its main metabolites in plasma after a simple liquid-liquid extraction.